[Histonet] oysters larvae again

Monfils, Paul PMonfils <@t> Lifespan.org
Tue Mar 21 10:23:04 CST 2006

I don't have a background in ISH, so I can't offer an opinion on the best
fixative for that purpose.  

Regarding your second question, while I'm not familiar with either fixative,
I can see that the Davidson fixative, containing 10% acetic acid and no
buffers, would decalcify (dissolve) larval oyster shells.  But the MEMPFA-T
appears to be made up in pH 7.4 buffer, in which case I don't see how it
would decalcify. In any case, I am wondering why you are using a
formaldehyde-based fixative for TEM?  Ordinarily a glutaraldehyde-based
fixative would be preferable for TEM.

> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of
> Anne-Sophie MARTINEZ
> Sent: 	Tuesday, March 21, 2006 5:13 AM
> To: 	histonet <@t> lists.utsouthwestern.edu
> Subject: 	[Histonet] oysters larvae again
> Hello,
> (1) I plan to fix oysters larvae/juveniles (4 month) for in toto ISH. 
> Which fixative is the best : Davidson (10% formaldehyde 37-40%, 30% 
> ethanol 95%, 60% seawater, 10% acetic acid) or MEMPFA-T (0,1 M Mops pH 
> 7,4; 2 mM EGTA; 1 mM MgSO4; 4% paraformaldehyde; 0,1% Tween 20)?
> (2) Would it be possible to use the MEMPFA-T to fix and "decalcify" the 
> shelves of the juveniles for TEM (embedding in type lowyril, unicril, 
> LR-white resins) instead of classical fixative (4% PFA) and then EDTA?
> Thank you very much again.
> Anne-Sophie
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